Removal of urine from the kidney is a vital process. Perturbations in urine flow can cause detrimental changes that manifest in the kidney itself. In the renal pelvis, peristaltic contractions of smooth muscle generate oscillatory forces that expel urine from the kidneys and into the ureters. Despite its importance, little is known about the mechanisms underlying the spontaneous nature of this process. A specialized smooth muscle cell has been hypothesized to be the putative pacemaker, but a unique histochemical marker for isolating this important cell type is not known. In this thesis, we have developed and validated a variety of fluorescent imaging approaches to study the renal pelvis at the inter- and intracellular levels. In addition to discovering a novel marker for pacemaker cells, we also concretely identified two novel interstitial cell populations that may have roles in modulating contractile activity. Finally, we determined that the Ca2+-activated chloride channel, Ano1, may be critical for ensuring rhythmic patterns of peristalsis. Taken together, this work addresses a major gap in the field and provides a novel toolkit that will enable future studies of cell type-specific contributions to renal pelvis function in health and disease.